Combination Therapies Using PRMT5 Inhibitors for the Treatment of Cancer

ABSTRACT

This disclosure relates to methods of treating cancer. This disclosure further relates to treating cancer in a subject with compounds that are inhibitors of PRMT5, particularly in combination with KRASG12C inhibitors.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from U.S. Provisional Application No.63/172,639, filed Apr. 8, 2021, and U.S. Provisional Application No.63/252,998, filed Oct. 6, 2021, the disclosure of each of which ishereby incorporated by reference in its entirety.

BACKGROUND OF THE DISCLOSURE Field of the Disclosure

This disclosure relates to methods of treating cancer. This disclosurefurther relates to treating cancer in a subject with compounds that areinhibitors of protein arginine N-methyl transferase 5 (PRMT5),particularly in combination with Kirsten rat sarcoma viral oncogenehomolog (KRAS) glycine-to-cysteine (G12C) (KRAS^(G12C)) inhibitors.

Description of Related Art

PRMT5 is a type II arginine methyltransferase that catalyzes thetransfer of a methyl group from S-adenosyl-L-methionine (SAM) to anomega-nitrogen of the guanidino function of protein L-arginine residues(omega-monomethylation) and the transfer of a second methyl group to theother omega-nitrogen, yielding symmetric dimethylarginine (sDMA). PRMT5forms a complex with methylosome protein 50 (MEP50), which is requiredfor substrate recognition and orientation and is also required forPRMT5-catalyzed histone 2A and histone 4 methyltransferase activity(e.g., see Ho et al. (2013) PLoS ONE 8(2): e57008).

Homozygous deletions of p16/CDKN2a are prevalent in cancer and thesemutations commonly involve the co-deletion of adjacent genes, includingthe gene encoding methylthioadenosine phosphorylase (MTAP). It isestimated that approximately 15% of all human cancers have a homozygousdeletion of the MTAP gene (e.g., see Firestone & Schramm (2017) J. Am.Chem Soc. 139(39):13754-13760).

Cells lacking MTAP activity have elevated levels of the MTAP substrate,methylthioadenosine (MTA), which is a potent inhibitor of PRMT5.Inhibition of PRMT5 activity results in reduced methylation activity andincreased sensitivity of cellular proliferation to PRMT5 depletion orloss of activity. Hence, the loss of MTAP activity reduces methylationactivity of PRMT5 making the cells selectively dependent on PRMT5activity.

Despite importance of PRMT5 on cell viability and its prevalence incancers, effective therapies that inhibit PRMT5 have been elusive. Thus,there remains a need to develop new PRMT5 inhibitor therapies to treatwide range of cancers.

SUMMARY OF THE DISCLOSURE

One aspect of the disclosure provides methods for treating cancer in asubject. Such methods include administering to the subject atherapeutically effective amount of a KRAS^(G12C) inhibitor and atherapeutically effective amount of a PRMT5 inhibitor.

Also provided herein is a method for treating cancer in a subject inneed thereof. Such methods include determining that the cancer isassociated with MTAP homozygous deletion (e.g., an MTAP-associatedcancer). These methods optionally further include determining that thecancer is associated with KRAS^(G12C) mutation. Such methods furtherinclude administering to the subject a therapeutically effective amountof a KRAS^(G12C) inhibitor and a therapeutically effective amount of aPRMT5 inhibitor.

These and other features and advantages of the present invention will bemore fully understood from the following detailed description takentogether with the accompanying claims. It is noted that the scope of theclaims is defined by the recitations therein and not by the specificdiscussion of features and advantages set forth in the presentdescription.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are included to provide a furtherunderstanding of the methods of the disclosure, and are incorporated inand constitute a part of this specification. The drawings illustrate oneor more embodiment(s) of the disclosure and, together with thedescription, serve to explain the principles and operation of thedisclosure.

FIG. 1 illustrates the results of the methods of Example 1 in theKRAS^(G12C) and CDKN2A/MTAP^(DEL) lung tumor xenograft LU99 model grownin immunodeficient mice. The PRMT5 inhibitor used in this method wasMRTX9768 administered at 100 mg/kg twice a day (BID), and theKRAS^(G12C) inhibitor was MRTX849 administered at 30 mg/kg once a day(QD). Average tumor volume±standard error is plotted of the mean atstudy day as indicated.

FIG. 2 illustrates the results of the methods of Example 2 in theKRAS^(G12C) and CDKN2A/MTAP^(DEL) lung tumor xenograft LU99 model grownin immunodeficient mice. The PRMT5 inhibitor used in this method wasMRTX7477 administered at 200 mg/kg BID, and the KRAS^(G12C) inhibitorwas MRTX849 administered at 30 mg/kg QD. Average tumor volume±standarderror is plotted of the mean at study day as indicated.

FIG. 3 illustrates the results of the methods of Example 3 in LU99PRMT5-041 lung tumor xenograft model. The PRMT5 inhibitor used in thismethod was MRTX1719 administered at 50 mg/kg QD, and theKRAS^(G12C)inhibitor was MRTX849 administered at 30 mg/kg QD. Averagetumor volume±standard error is plotted of the mean at study day asindicated.

FIG. 4 illustrates the results of the methods of Example 4 in SW1573PRMT5-044 lung tumor xenograft model. The PRMT5 inhibitor used in thismethod was MRTX1719 administered at 50 mg/kg QD, and the KRAS^(G12C)inhibitor was MRTX849 administered at 100 mg/kg QD. Average tumorvolume±standard error is plotted of the mean at study day as indicated.

DETAILED DESCRIPTION OF THE DISCLOSURE

Before the disclosed processes and materials are described, it is to beunderstood that the aspects described herein are not limited to specificembodiments, and as such can, of course, vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular aspects only and, unless specifically definedherein, is not intended to be limiting.

In view of the present disclosure, the methods and compositionsdescribed herein can be configured by the person of ordinary skill inthe art to meet the desired need. The present disclosure providesimprovements in treating cancer in a subject. As used herein, the terms“subject” or “patient” are used interchangeably, refers to any animal,including mammals, and most preferably humans.

The methods provided herein may be used for the treatment of a widevariety of cancer including tumors such as lung, prostate, breast,brain, skin, cervical carcinomas, testicular carcinomas, etc. Moreparticularly, cancers that may be treated by the compositions andmethods of the invention include, but are not limited to tumor typessuch as astrocytic, breast, cervical, colorectal, endometrial,esophageal, gastric, head and neck, hepatocellular, laryngeal, lung,oral, ovarian, prostate and thyroid carcinomas and sarcomas. Morespecifically, these compounds can be used to treat: Cardiac: sarcoma(angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma,rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma(squamous cell, undifferentiated small cell, undifferentiated largecell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchialadenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma,leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma,leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma,glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel(adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor(nephroblastoma), lymphoma, leukemia), bladder and urethra (squamouscell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone:osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumorchordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma (serouscystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiedcarcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma(embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic:blood (myeloid leukemia (acute and chronic), acute lymphoblasticleukemia, chronic lymphocytic leukemia, myeloproliferative diseases,multiple myeloma, myelodysplastic syndrome), Hodgkin's disease,non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma,basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, molesdysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis;and Adrenal glands: neuroblastoma.

In certain embodiments of the methods of the disclosure, the cancer is aMTAP-associated cancer. For example, in certain embodiments, the cancercomprises MTAP gene homozygous deletion (MTAPDEL). The subject may beidentified or diagnosed as having MTAP-associated cancer where, forexample, MTAPDEL is determined using a suitable assay or a kit.Alternatively, the subject is suspected of having MTAP-associated canceror the subject has a clinical record indicating that the subject hasMTAP-associated cancer.

In certain embodiments of the methods of the disclosure, the cancercomprises a KRAS^(G12C) gene mutation. The subject may be identified ordiagnosed as having KRAS^(G12C) cancer where KRAS^(G12C) mutation isdetermined using a suitable assay or a kit. Alternatively, the subjectis suspected of having the KRAS^(G12C) cancer or the subject has aclinical record indicating that the subject has the KRAS^(G12C) cancer.

In certain embodiments of the methods of the disclosure, the cancer mayfurther comprise a cyclin-dependent kinase inhibitor 2A (CDKN2A) genehomozygous deletion (CDKN2A^(DEL)). The subject may be identified ordiagnosed as having CDKN2A^(DEL) where the deletion is determined usinga suitable assay or a kit. Alternatively, the subject is suspected ofhaving the CDKN2A^(DEL) cancer, or the subject has a clinical recordindicating that the subject has the CDKN2A^(DEL) cancer.

In some embodiments of any of the methods or uses described herein, anassay is used to determine whether the patient has MTAP^(DEL) and/orKRAS^(G12C) and/or CDKN2A^(DEL) using a sample (e.g., a biologicalsample or a biopsy sample such as a paraffin-embedded biopsy sample)from a subject. Such assay includes, but is not limited to, nextgeneration sequencing, immunohistochemistry, fluorescence microscopy,break apart FISFI analysis, Southern blotting. Western blotting, FACSanalysis, Northern blotting, and PCR-based amplification (e.g., RT-PCRand quantitative real-time RT-PCR). As is well known in the art, theassays are typically performed, e.g., with at least one labelled nucleicacid probe or at least one labelled antibody or antigen-binding fragmentthereof.

In certain embodiments, the cancer in the methods of the disclosure isselected from lung cancer, pancreatic cancer, colon cancer, head andneck cancer, bladder cancer, esophageal cancer, lymphoma, stomachcancer, skin cancer, breast cancer, and brain cancer.

In certain embodiments, the cancer in the methods of the disclosure isselected from lung cancer, pancreatic cancer, colon cancer, head andneck cancer, esophageal cancer, and melanoma.

In certain embodiments, the cancer in the methods of the disclosure isselected from lung cancer (e.g., mesothelioma or non-small cell lungcancer (NSCLC) including adenocarcinoma and squamous cell), pancreaticcancer, colon cancer, head and neck cancer (such as squamous cellcarcinoma (HNSCC)), bladder cancer, esophageal cancer, lymphoma (e.g.,diffuse large B-cell lymphoma), stomach cancer, melanoma, breast cancer,and brain cancer (e.g., glioblastoma multiforme and glioma).

In certain embodiments, the cancer in the methods of the disclosure isselected from lung cancer (e.g., mesothelioma or NSCLC, includingadenocarcinoma and squamous cell), pancreatic cancer, colon cancer, headand neck cancer (e.g. squamous cell carcinoma (HNSCC)), esophagealcancer, and melanoma.

In certain embodiments, the cancer in the methods of the disclosure isselected from mesothelioma, NSCLC (e.g., adenocarcinoma and squamouscell), pancreatic cancer, HNSCC, and colon cancer.

In one embodiment of the methods of the disclosure, the cancer is lungcancer. For example, the lung cancer may be NSCLC (e.g., adenocarcinomaand squamous cell) or mesothelioma. In certain embodiment, the cancer isNSCLC.

In one embodiment of the methods of the disclosure, the cancer ispancreatic cancer.

In one embodiment of the methods of the disclosure, the cancer is coloncancer.

As provided above, the KRAS^(G12C) inhibitor is administered in themethods of the disclosure. As used herein, a “KRAS^(G12C) inhibitor”refers to compounds capable of negatively modulating or inhibiting allor a portion of the enzymatic activity of KRAS^(G12C). The KRAS^(G12C)inhibitors of the present disclosure interact with and/or irreversiblybind to KRAS^(G12C) (e.g., by forming a covalent adduct with thesulfhydryl side chain of the cysteine residue at position 12) resultingin the inhibition of the enzymatic activity of KRAS^(G12C).

In certain embodiments, the KRAS^(G12C) inhibitor is selected fromadagrasib (shown below, also known as MRTX849, Mirati Therapeutics,Inc., San Diego, Calif.), sotorasib (also known as AMG510, Amgen Inc.,Thousand Oaks, Calif.), JNJ-74699157 (also known as ARS-3248, JanssenResearch & Development, LLC, Raritan, N.J.), GDC-6036 (Roche, Basel,Switzerland) , LY3499446 (Eli Lilly and Company, Indianapolis, Ind.),JDQ443 (Novartis Pharmaceuticals, Basel, Switzerland), D-1553(InventisBio Inc., Shanghai, China), and combinations thereof.

In one embodiment of the methods of the disclosure, the KRAS^(G12C)inhibitor is adagrasib or sotorasib. In one embodiment, the KRAS^(G12C)inhibitor is sotorasib.

In one embodiment, the KRAS^(G12C) inhibitor is adagrasib. Adagrasib hasthe following structure:

In certain embodiments, the KRAS^(G12C) inhibitor of the disclosure isany one of the KRAS^(G12C) inhibitors disclosed in International patentpublication Nos. WO 2017/201161 A1, published 23 Nov. 2017, WO2019/099524 A1, published 23 May 2019, WO 2019/217307 A1, published 14Nov. 2019, WO 2020/047192 A1, published 5 Mar. 2020, WO 2020/101736 A1,published 22 May 2020, or WO 2020/146613 A1, published 16 Jul. 2020, allincorporated by reference in their entirety.

As provided above, the PRMT5 inhibitor is also administered in themethods of the disclosure. A “PRMT5 inhibitor” as used herein refers tocompounds of the disclosure as described herein. These compounds arecapable of negatively modulating or inhibiting all or a portion of theenzymatic activity of the PRMT5, particularly, in the presence of boundMTA in vitro or in vivo or in cells expressing elevated levels of MTA.In certain embodiments, the PRMT5 inhibitor is a MTA-cooperative PRMT5inhibitor.

In certain embodiments, the PRMT5 inhibitor of the disclosure is any oneof the PRMT5 inhibitors disclosed in International patent publicationNo. WO 2021/050915 A1, published 18 Mar. 2021, incorporated by referencein its entirety.

In certain other embodiments, the PRMT5 inhibitor of the disclosure isany one of the PRMT5 inhibitors disclosed in U.S. provisionalapplication No. 63/200,521, filed 11 Mar. 2021, incorporated byreference in its entirety.

For example, the PRMT5 inhibitor in the methods of the disclosure asdescribed herein is a compound of Formula IIA, IIB or IIC (Embodiment1):

or a pharmaceutically acceptable salt thereof, wherein:

A is CR⁹ or N;

D is (C(R⁹)₂)₁₋₂-NH₂,

or D is

where the methylene is bonded to E where E is C;

E is C, CR⁹ or N;

each L is independently a bond or C₁-C₃ alkylene;

W is CR⁹ or N;

each X is independently a bond, O, S, —NR⁴— or —NR⁴C(O)—;each Z is independently a bond, —SO—, —SO₂—, —CH(OH)— or —C(O)—;each R² is independently hydroxy, halogen, cyano, cyanomethyl, —(NR⁴)₂,hydroxyalkyl, alkoxy, —SO₂C₁-C₃alkyl, —X-arC₁-C₃alkyl, heteroalkyl,C₂-C₄ alkynyl, —X-haloalkyl, —X—C₁-C₅ alkyl, —Z—C₁-C₅ alkyl,heterocyclyl, —X—L-cycloalkyl, —Z-cycloalkyl, —X-aryl, —Z-aryl, or—X-heteroaryl, wherein the heterocyclyl, the cycloalkyl, the aryl andthe heteroaryl are optionally substituted with one or more R⁵;each R⁴ is independently hydrogen or C₁-C₃ alkyl;each R⁵ is independently cyano, oxo, halogen, C₁-C₃ alkyl, hydroxyalkyl,hydroxy, alkoxy, alkoxy-C₁-C₃ alkyl, —X-haloalkyl, —Z-cycloalkyl,—X-arC₁-C₃alkyl, —X-arC₁-C₃alkyl substituted with cyano, —X—L-cycloalkyloptionally substituted with C₁-C₃ alkyl or oxo, —X—L-heteroaryloptionally substituted with one or more C₁-C₃ alkyl or oxo,—X—L-heterocyclyl optionally substituted with one or more C₁-C₃ alkyl oroxo, or —X-aryl;R⁶ is hydrogen, halogen, C₁-C₃ alkyl, haloalkyl, hydroxy, alkoxy, C₁-C₃alkyl-alkoxy, N(R⁹)₂, NR⁹C(O)R⁹, C(O)R⁹, oxetane and THF;R⁷ is H or C₁-C₃ alkyl optionally substituted with one or more halogen;R⁸ is H or C₁-C₃ alkyl; andeach R⁹ is independently H or C₁-C₃ alkyl, halogen or haloalkyl.

Embodiment 2 provides the PRMT5 inhibitor in the methods of thedisclosure as a compound of Formula IIA:

Embodiment 3 provides the PRMT5 inhibitor in the methods of thedisclosure as a compound of Formula IIB:

Embodiment 4 provides the PRMT5 inhibitor in the methods of thedisclosure as a compound of Formula IIC:

Embodiment 5 provides the method of any of embodiments 1-4, wherein W isCR⁹.

Embodiment 6 provides the method of any of embodiments 1-4, wherein A isCR⁹.

Embodiment 7 provides the method of any of embodiments 1-4, wherein E isN.

Embodiment 8 provides the method of any of embodiments 1-7, wherein W isCR⁹, A is CR⁹ and E is N.

Embodiment 9 provides the method of any of embodiments 1-8, wherein R²is selected from: benzothiophene, naphthalene, quinoline, chromane,isochromane, dihydrobenzodioxine, indolazine, tetrahydroindolazine,dihydroisobenzofuran, benzene, isoquinolinone, benzodioxone,thienopyridine, tetrahydroindolone, indolizine, dihydroindolizinone,imadazopyridinone, thienopyrimidine, thiophene, pyrrolopyrimidinone,thiazolopyridinone, dihydropyrrolizine, isoindalone andtetrahydroisoquinoline.

Embodiment 10 provides the method of any of embodiments 1-8, whereineach R⁵ is independently cyano, oxo, halogen, C1-C3 alkyl, hydroxy,hydroxyalkyl, alkoxy-C1-C3alkyl, —X—L-heterocyclyl optionallysubstituted with one or more C1-C3alkyl or oxo, —X—L-cycloalkyloptionally substituted with C1-C3 alkyl or oxo.

Embodiment 11 provides the method of any of embodiments 1-8, wherein R⁶is selected from hydrogen, hydroxy, chlorine, —NHC(O)CH₃, —C(O)CF₂H,—NH₂, —CF₂, —CH₃, —O—CH₂CH₃, —CH₂—CH₂—O—CH₃, oxetane and THF.

Embodiment 12 provides the method of any of embodiments 1-11, where oneof L, X and Z is a bond.

Embodiment 13 provides the method of embodiment 12, wherein all of L, Xand Z are bonds.

One aspect of the disclosure provides the method wherein the PRMT5inhibitor is a compound of the formula (IIIC) (Embodiment 14):

or a pharmaceutically acceptable salt thereof, wherein

A is CR⁹ or N;

D is —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl;G, Q, J and U are independently selected from C(H), C(R⁵), and N,provided only one or two of G, Q, J, and U can be N;

-   -   each R⁵ is independently hydroxy, halogen, C₁-C₆ alkyl, C₁-C₆        haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, C₃-C₆ cycloalkoxy,        C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃ alkoxyC₁-C₃        alkyl;        R⁶ is hydrogen, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, hydroxy,        C₁-C₆ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,        —C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶,    -   where each R⁹ is independently H or C₁-C₃ alkyl, R¹⁵ is hydrogen        or methyl, and R¹⁶ is C₁-C₃ alkyl; and        R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.

Embodiment 15 provides the method according to embodiment 14, wherein Ais CH.

Embodiment 16 provides the method according to embodiment 14 or 15,wherein W is N.

Embodiment 17 provides the method according to embodiment 14 or 15,wherein W is CH.

Embodiment 18 provides the method according to any of embodiments 14-17,wherein D is —CH₂—NH₂.

Embodiment 19 provides the method of the disclosure wherein the PRMT5inhibitor is a compound according to embodiment 14 of the formula:

Embodiment 20 provides the method according to any of embodiments 14-19,wherein R⁶ is hydrogen, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, hydroxy,C₁-C₆ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,—C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 21 provides the method according to any of embodiments 14-19,wherein R⁶ is hydrogen, halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy,C₁-C₃ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,—C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 22 provides the method according to any of embodiments 14-19,wherein R⁶ is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl,hydroxy, methoxy, ethoxy, (methoxy) methyl, (ethoxy) methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl,—NH₂, or —NH(CO)CH₃.

Embodiment 23 provides the method according to any of embodiments 14-19,wherein R⁶ is halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, hydroxy, C₁-C₆alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 24 provides the method according to any of embodiments 14-19,wherein R⁶ is halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy, C₁-C₃alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)-C₁-C₃haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 25 provides the method according to any of embodiments 14-19,wherein R⁶ is chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy,methoxy, ethoxy, (methoxy) methyl, (ethoxy) methyl, (methoxy)ethyl,(ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH₂,or —NH(CO)CH₃.

Embodiment 26 provides the method according to any of embodiments 23-25,wherein each G, Q, J and U is independently C(H).

Embodiment 27 provides the method according to any of embodiments 23-25,wherein G, Q, J and U are independently selected from C(H) and C(R⁵).

Embodiment 28 provides the method according to any of embodiments 23-25,wherein G, Q, J and U are independently selected from C(H) and N.

Embodiment 29 provides the method according to any of embodiments 14-19,wherein

-   -   R⁶ is hydrogen;    -   at least one of G, Q, J, and U is C(R⁵), and the remaining G, Q,        J, and U are independently selected from C(H), C(R⁵) and N,        wherein each R⁵ is independently hydroxy, halogen, C₁-C₆ alkyl,        C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, C₃-C₆        cycloalkoxy, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃        alkoxyC₁-C₃ alkyl.

Embodiment 30 provides the method according to embodiment 29, whereinone or two of G, Q, J and U is N.

Embodiment 31 provides the method according to any of embodiments 14-19,wherein

-   -   R⁶ is hydrogen;    -   at least one of G, Q, J, and U is C(R⁵), and the remaining G, Q,        J, and U are independently selected from C(H) and C(R⁵), wherein        each R⁵ is independently hydroxy, halogen, C₁-C₆ alkyl, C₁-C₆        haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, C₃-C₆ cycloalkoxy,        C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃ alkoxyC₁-C₃        alkyl.

Embodiment 32 provides the method according to embodiment 31, wherein atleast one of G, Q, J, and U is C(R⁵), and the remaining G, Q, J, and Uare independently C(H); for example only one of G, Q, J, and U is C(R⁵).

Embodiment 33 provides the method according to embodiment 31, whereintwo of G, Q, J, and U is C(R⁵), and the remaining G, Q, J, and U areindependently C(H).

Embodiment 34 provides the method according to embodiment 31, whereinthree of G, Q, J, and U is C(R⁵), and the remaining G, Q, J, and U isC(H).

Embodiment 35 provides the method according to any of embodiments 14-19,wherein G, Q, J, and U together with the thiophene to which they areattached form:

Embodiment 36 provides the method according to embodiment 35, wherein G,Q, J, and U together with the thiophene ring to which they are attachedform a benzo[b]thiophene.

Embodiment 37 provides the method according to any one of embodiments14-36, wherein R⁵, if present, is hydroxy, halogen, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ cycloalkoxy, C₃-C₆cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃ alkoxyC₁-C₃ alkyl.

Embodiment 38 provides the method according to any one of embodiments14-36, wherein R⁵, if present, is hydroxy, halogen, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ heterocycloalkyl, orC₁-C₃ alkoxyC₁-C₃ alkyl.

Embodiment 39 provides the method according to any one of embodiments14-36, wherein R⁵, if present, is hydroxy, chloro, fluoro, methyl,ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl,(methoxy) methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

Embodiment 40 provides the method according to any one of embodiments14-39, wherein R⁷ is methyl.

Embodiment 41 provides the method according to any one of embodiments14-39, wherein R⁷ is ethyl.

Embodiment 42 provides the method according to any one of embodiments14-39, wherein R⁷ is propyl (e.g., isopropyl).

Embodiment 43 provides the method according to any one of embodiments14-39, wherein R⁷ is difluoromethyl or trifluoromethyl.

Embodiment 44 provides the method according to embodiment 14, whereinthe PRMT5 inhibitor is of the formula:

whereinG, Q, J, and U together with the thiophene to which they are attachedform:

where each R⁵ is independently hydroxy, halogen, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ heterocycloalkyl, orC₁-C₃ alkoxyC₁-C₃ alkyl; and

-   -   R⁶ is hydrogen, halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy,        C₁-C₃ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,        —C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 45 provides the method according to embodiment 14, whereinthe PRMT5 inhibitor is of the formula:

whereinG, Q, J, and U together with the thiophene to which they are attachedform:

where each R⁵ is independently hydroxy, halogen, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ heterocycloalkyl, orC₁-C₃ alkoxyC₁-C₃ alkyl; andR⁶ is halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy, C₁-C₃ alkoxy,C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃ haloalkyl,—N(R⁹)₂, or —NR¹⁵(CO)R¹⁶.

Embodiment 46 provides the method according to embodiment 14, whereinthe PRMT5 inhibitor is of the formula:

whereinG, Q, J, and U together with the thiophene to which they are attachedform:

where each R⁵ is independently hydroxy, halogen, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ heterocycloalkyl, orC₁-C₃ alkoxyC₁-C₃ alkyl.

Embodiment 47 provides the method of the disclosure wherein the PRMT5inhibitor is a compound of the formula (IIIB):

or a pharmaceutically acceptable salt thereof, wherein

A is CR⁹ or N;

D is —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl;R⁵¹ is hydrogen, fluoro, chloro, or methyl, or R⁵¹ and R⁵² together withatoms to which they are attached form a C₄-C₆ heterocycloalkyl (e.g,hydrofuranyl);R⁵² is fluoro, chloro, or methyl, or R⁵² and R⁵³ together with atoms towhich they are attached form a phenyl;R⁵³ is hydrogen, fluoro, chloro, or methyl;R⁵⁴ is hydrogen, halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy;L⁵ is —O— or —CH₂—;R⁶ is hydrogen, halogen, C₁-C₆ alkyl, hydroxy, C₁-C₆ alkoxy, C₁-C₃alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃ haloalkyl, or—NR¹⁵(CO)R¹⁶, where R¹⁵ is hydrogen or methyl, and R¹⁶ is C₁-C₃ alkyl;R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.

Embodiment 48 provides the method according to embodiment 47, wherein:

-   -   A is —CH or —CCH₃;    -   D is —CH₂—NH₂;    -   W is —CH, —CCH₃, or N;    -   R⁵¹, R⁵², R⁵³, and R⁵⁴ are each independently selected from        hydrogen, fluoro, chloro, or methyl;    -   L⁵ is —O—;    -   R⁶ is hydrogen, fluoro, chloro, or methyl; and    -   R⁷ is C₁-C₂ alkyl or C₁-C₂ haloalkyl.

Embodiment 49 provides the method according to embodiment 47 orembodiment 48, wherein:

-   -   A and W are —CH;    -   D is —CH₂—NH₂;    -   R⁵¹, R⁵², and R⁵³ are each independently selected from hydrogen,        fluoro, chloro, and methyl;    -   R⁵⁴ is hydrogen;    -   L⁵ is —O—;    -   R⁶ is hydrogen; and    -   R⁷ is methyl.

Embodiment 50 provides the method according to any of embodiments 47-49,wherein:

-   -   A and W are —CH;    -   D is —CH₂—NH₂;    -   R⁵¹ and R⁵² are each independently selected from fluoro, chloro,        and methyl;    -   R⁵³ and R⁵⁴ are hydrogen;    -   L⁵ is —O—;    -   R⁶ is hydrogen; and    -   R⁷ is methyl.

Embodiment 51 provides the method according to embodiment 47, wherein Ais CH.

Embodiment 52 provides the method according to embodiment 47 or 48,wherein W is N.

Embodiment 53 provides the method according to embodiment 47 or 48,wherein W is CH.

Embodiment 54 provides the method according to any of embodiments 47-50,wherein D is —CH₂—NH₂.

Embodiment 55 provides the method according to any of embodiments 47-51,wherein R⁵⁴ is hydrogen or methyl.

Embodiment 56 provides the method according to any of embodiments 47-51,wherein R⁵⁴ is hydrogen.

Embodiment 57 provides the method according to any of embodiments 47-51,wherein R⁵⁴ is methyl.

Embodiment 58 provides the method according to embodiment 47, where thePRMT5 inhibitor is of the formula:

such as e.g.,

Embodiment 59 provides the method according to any of embodiments 47-55,wherein L⁵ is —CH₂—.

Embodiment 60 provides the method according to any of embodiments 47-55,wherein L⁵ is —O—.

Embodiment 61 provides the method according to any of embodiments 47-57,wherein R⁶ is hydrogen, halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy,C₁-C₃ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,—C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶; for example, wherein R⁶is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy,methoxy, ethoxy, (methoxy)methyl, (ethoxy) methyl, (methoxy)ethyl,(ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH₂,or —NH(CO)CH₃.

Embodiment 62 provides the method according to any of embodiments 47-57,wherein R⁶ is hydrogen, halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; forexample, R⁶ is hydrogen, halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy.

Embodiment 63 provides the method according to any of embodiments 47-57,wherein R⁶ is hydrogen, chloro, fluoro, methyl, ethyl, methoxy, orethoxy.

Embodiment 64 provides the method according to any of embodiments 47-57,wherein R⁶ is halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy, C₁-C₃alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶; for example, wherein R⁶ is chloro,fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy,(methoxy) methyl, (ethoxy) methyl, (methoxy) ethyl, (ethoxy)ethyl,oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH₂, or —NH(CO)CH₃.

Embodiment 65 provides the method according to any of embodiments 47-57,wherein R⁶ is halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; for example, R⁶ ishalogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy.

Embodiment 66 provides the method according to any of embodiments 47-57,wherein R⁶ is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

Embodiment 67 provides the method according to any one of embodiments47-63, wherein R⁷ is methyl.

Embodiment 68 provides the method according to any one of embodiments47-63, wherein R⁷ is ethyl.

Embodiment 69 provides the method according to any one of embodiments47-63, wherein R⁷ is propyl (e.g., isopropyl).

Embodiment 70 provides the method according to any one of embodiments47-63, wherein R⁷ is difluoromethyl or trifluoromethyl.

Embodiment 71 provides the method according to any of embodiments 47-67,wherein R⁵³ is hydrogen or methoxy; or wherein R⁵³ is hydrogen.

Embodiment 72 provides the method according to embodiment 47, where thePRMT5 inhibitor is of the formula:

Embodiment 73 provides the method according to any one of embodiments47-69, wherein R⁵² is fluoro, and R⁵¹ is hydrogen, fluoro, chloro, ormethyl.

Embodiment 74 provides the method according to any one of embodiments47-69, wherein R⁵² is fluoro, and R⁵¹ is chloro.

Embodiment 75 provides the method according to any one of embodiments47-69, wherein R⁵² is fluoro, and R⁵¹ is methyl or hydrogen (forexample, R⁵² is fluoro and R⁵¹ is methyl; or R⁵² is fluoro and R⁵¹ ishydrogen).

Embodiment 76 provides the method according to any one of embodiments47-69, wherein R⁵¹ and R⁵² together with atoms to which they areattached form a hydrofuranyl

Embodiment 77 provides the method according to any one of embodiments47-76, wherein the PRMT5 inhibitor is

Embodiment 78 provides the method according to any one of embodiments47-77, wherein the PRMT5 inhibitor is

One aspect of the disclosure provides the method wherein the PRMT5inhibitor is a compound of the formula (IIIA) (Embodiment 79):

or a pharmaceutically acceptable salt thereof, wherein A is CR⁹ or N;D is —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl; R² is

where R⁵⁶ is hydrogen, fluoro, chloro, or methyl,G, Q, J and U are independently selected from C(H), C(R⁵), and N,provided only one or two of G, Q, J, and U can be N;

-   -   each R⁵ is independently hydroxy, halogen, C₁-C₆ alkyl, C₁-C₆        haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, C₃-C₆ cycloalkoxy,        C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃ alkoxyC₁-C₃        alkyl;        R⁶ is hydrogen, halogen, C₁-C₆ alkyl, hydroxy, C₁-C₆ alkoxy,        C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃        haloalkyl, or —NR¹⁵(CO)R¹⁶, where R¹⁵ is hydrogen or methyl, and        R¹⁶ is C₁-C₃alkyl; and        R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.

One aspect of the disclosure provides the method wherein the PRMT5inhibitor is a compound of the formula (IIIA) (Embodiment 80):

or a pharmaceutically acceptable salt thereof, wherein

A is CR⁹ or N;

D is —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl;

R² is

where R⁵⁶ is hydrogen, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆alkoxy, or C₁-C₆ haloalkoxy;R⁶ is hydrogen, halogen, C₁-C₆ alkyl, hydroxy, C₁-C₆ alkoxy, C₁-C₃alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃ haloalkyl, or—NR¹⁵(CO)R¹⁶, where R¹⁵ is hydrogen or methyl, and R¹⁶ is C₁-C₃alkyl;andR⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.

Embodiment 81 provides the method according to embodiment 79 or 80,wherein A is CH.

Embodiment 82 provides the method according to embodiment 79 or 80,wherein W is N.

Embodiment 83 provides the method according to embodiment 79 or 80,wherein W is CH.

Embodiment 84 provides the method according to any of embodiments 79 or80, wherein D is —CH₂—NH₂.

Embodiment 85 provides the method according to embodiment 79 or 80,which is of the formula:

Embodiment 86 provides the method according to embodiment 79 or 81-85,wherein R² is

Embodiment 87 provides the method according to embodiment 86, wherein G,Q, J and U are independently selected from C(H) and C(R⁵).

Embodiment 88 provides the method according to embodiment 86, wherein G,Q, J and U are independently C(H).

Embodiment 89 provides the method according to embodiment 86, wherein atleast one of G, Q, J, and U is C(R⁵), and the remaining G, Q, J, and Uare independently C(H); for example only one of G, Q, J, and U is C(R⁵).

Embodiment 90 provides the method according to embodiment 86, wherein Uis N, and G, Q, and J are independently selected from C(H) and C(R⁵).

Embodiment 91 provides the method according to embodiment 86, wherein Gis N, and Q, J, and U are independently selected from C(H) and C(R⁵).

Embodiment 92 provides the method according to any one of embodiments 79or 81-91, wherein R⁵, if present, is hydroxy, halogen, C₁-C₃ alkyl,C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ cycloalkoxy,C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃ alkoxyC₁-C₃ alkyl.

Embodiment 93 provides the method according to any one of embodiments 79or 81-91, wherein R⁵, if present, is hydroxy, halogen, C₁-C₃ alkyl,C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, C₃-C₆ heterocycloalkyl,or C₁-C₃ alkoxyC₁-C₃ alkyl.

Embodiment 94 provides the method according to any one of embodiments 79or 81-91, wherein R⁵, if present, is hydroxy, chloro, fluoro, methyl,ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl,(methoxy) methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

Embodiment 95 provides the method according to any one of embodiments 79or 81-91, wherein R⁵, if present, is halogen, C₁-C₆ alkyl, or C₁-C₆alkoxy; for example, R⁶ is halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy.

Embodiment 96 provides the method according to any one of embodiments 79or 81-91, wherein R⁵, if present, is chloro, fluoro, methyl, ethyl,methoxy, or ethoxy.

Embodiment 97 provides the method according to any one of embodiments 79or 81-91, wherein R⁵⁶ is fluoro, chloro, or methyl.

Embodiment 98 provides the method according to embodiment 80-85, whereinR² is

Embodiment 99 provides the method according to any of embodiments 80-85or 98, wherein R⁵⁶ is hydrogen, fluoro, chloro, or methyl.

Embodiment 100 provides the method according to any of embodiments79-99, wherein R⁶ is hydrogen, halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl,hydroxy, C₁-C₃ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,—C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶; for example, wherein R⁶is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy,methoxy, ethoxy, (methoxy)methyl, (ethoxy) methyl, (methoxy)ethyl,(ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH₂,or —NH(CO)CH₃.

Embodiment 101 provides the method according to any of embodiments79-99, wherein R⁶ is hydrogen, halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy;for example, R⁶ is hydrogen, halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy.

Embodiment 102 provides the method according to any of embodiments79-99, wherein R⁶ is hydrogen, chloro, fluoro, methyl, ethyl, methoxy,or ethoxy.

Embodiment 103 provides the method according to any of embodiments79-99, wherein R⁶ is halogen, C₁-C₃ alkyl, C₁-C₃ haloalkyl, hydroxy,C₁-C₃ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl,—C(O)-C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶; for example, wherein R⁶is chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy,ethoxy, (methoxy) methyl, (ethoxy) methyl, (methoxy) ethyl,(ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH₂,or —NH(CO)CH₃.

Embodiment 104 provides the method according to any of embodiments79-99, wherein R⁶ is halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; for example,R⁶ is halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy.

Embodiment 105 provides the method according to any of embodiments79-99, wherein R⁶ is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

Embodiment 106 provides the method according to any one of embodiments79-105, wherein R⁷ is methyl.

Embodiment 107 provides the method according to any one of embodiments79-105, wherein R⁷ is ethyl.

Embodiment 108 provides the method according to any one of embodiments79-105, wherein R⁷ is propyl (e.g., isopropyl).

Embodiment 109 provides the method according to any one of embodiments79-105, wherein R⁷ is difluoromethyl or trifluoromethyl.

In certain embodiments of the methods of the disclosure as describedherein, the PRMT5 inhibitor is:

In certain embodiments of the methods of the disclosure as describedherein, the PRMT5 inhibitor is:

In certain embodiments of the methods of the disclosure as describedherein, the PRMT5 inhibitor is:

In certain embodiments of the methods of the disclosure as describedherein, the PRMT5 inhibitor is:

The PRMT5 inhibitor of the disclosure and/or the KRAS^(G12C) inhibitorof the disclosure may be provided as a pharmaceutical compositioncomprising a therapeutically effective amount of such inhibitor and apharmaceutically acceptable carrier, excipient, and/or diluents. ThePRMT5 inhibitor of the disclosure and/or the KRAS^(G12C) inhibitor ofthe disclosure may be formulated by any method well known in the art andmay be prepared for administration by any route, including, withoutlimitation, parenteral, oral, sublingual, transdermal, topical,intranasal, intratracheal, or intrarectal. In certain embodiments, ThePRMT5 inhibitor of the disclosure and/or the KRAS^(G12C) inhibitor ofthe disclosure are administered intravenously in a hospital setting. Incertain other embodiments, administration may preferably be by the oralroute.

The characteristics of the carrier will depend on the route ofadministration. As used herein, the term “pharmaceutically acceptable”means a non-toxic material that is compatible with a biological systemsuch as a cell, cell culture, tissue, or organism, and that does notinterfere with the effectiveness of the biological activity of theactive ingredient(s). Thus, pharmaceutical compositions of thedisclosure may contain, in addition to the inhibitor, diluents, fillers,salts, buffers, stabilizers, solubilizers, and other materials wellknown in the art. The preparation of pharmaceutically acceptableformulations is described in, e.g., Remington's Pharmaceutical Sciences,18^(th) Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

The PRMT5 inhibitor and the KRAS^(G12C) inhibitor of the disclosure areadministered in a therapeutically effective amount. As used herein, thephrase “therapeutically effective amount” or “effective amount” refersto the amount of active agent that elicits the biological or medicinalresponse that is being sought in a tissue, system, subject or human by aresearcher, medical doctor or other clinician. In general, thetherapeutically effective amount is sufficient to deliver the biologicalor medicinal response to the subject without causing serious toxiceffects. A dose of the active agent may be in the range from about 0.01to 300 mg/kg per day, such as 0.1 to 100 mg/kg per day, more generally0.5 to about 25 mg/kg body weight of the recipient per day. A typicaltopical dosage will range from 0.01 to 3% wt/wt in a suitable carrier.

In certain embodiments of the methods of the disclosure, thetherapeutically effective amount of the PRMT5 inhibitor is in the rangeof about 0.01 to 300 mg/kg per day. For example, in certain embodiments,the therapeutically effective amount of the PRMT5 inhibitor is in therange of about 0.1 to 100 mg/kg per day, or 25 to 100 mg/kg per day, or50 to 100 mg/kg per day.

In certain embodiments, the therapeutically effective amount of thePRMT5 inhibitor is less than 1% of, e.g., less than 10%, or less than25%, or less than 50% of the clinically-established therapeutic amount(e.g., such as the amount required when the PRMT5 inhibitor isadministered by itself).

In certain embodiments of the methods of the disclosure, thetherapeutically effective amount of the KRAS^(G12C) inhibitor is in therange of about 0.01 to 300 mg/kg per day. For example, in certainembodiments, the therapeutically effective amount of the KRAS^(G12C)inhibitor is in the range of about 0.1 to 100 mg/kg per day, or 0.1 to50 mg/kg per day, or 10 to 100 mg/kg per day, or 10 to 50 mg/kg per day.

In certain embodiments, the therapeutically effective amount of theKRAS^(G12C) inhibitor is less than 1% of, e.g., less than 10%, or lessthan 25%, or less than 50% of the clinically-established therapeuticamount (e.g., such as the amount required when the KRAS^(G12C) inhibitoris administered by itself).

Combination therapy, in defining use of PRMT5 inhibitor and theKRAS^(G12C) inhibitor of the present disclosure, is intended to embraceadministration of each agent in a sequential manner in a regimen thatwill provide beneficial effects of the drug combination (e.g., the PRMT5inhibitor and the KRAS^(G12C) inhibitor of the disclosure can beformulated as separate compositions that are given sequentially), and isintended as well to embrace co-administration of these agents in asubstantially simultaneous manner, such as in a single dosage formhaving a fixed ratio of these active agents or in multiple or a separatedosage forms for each agent. The disclosure is not limited in thesequence of administration: the PRMT5 inhibitor of the disclosure may beadministered either prior to or after (i.e., sequentially), or at thesame time (i.e., simultaneously) as administration of the KRAS^(G12C)inhibitor of the disclosure.

The methods of disclosure are useful as a first-line treatment. Thus, incertain embodiments of the methods of the disclosure, the subject hasnot previously received another first-line of therapy.

The methods of disclosure are also useful as a first-line maintenance ora second-line treatment. Thus, in certain embodiments of the methods ofthe disclosure, the subject has previously completed another first-lineof therapy. For example, the methods of the disclosure, in certainembodiments, may provide a delay in progression and relapse of cancer insubjects that have previously completed another first-line chemotherapy.For example, in certain embodiments, the subject has previouslycompleted a platinum- and/or taxane-based chemotherapy (e.g.,carboplatin, cisplatin, oxaliplatin, paclitaxel, docetaxel, and thelike). In certain embodiments of the methods of the disclosure, thesubject has previously completed another first-line chemotherapy and isin partial response to such chemotherapy.

Definitions

For simplicity, chemical moieties are defined and referred to throughoutprimarily as univalent chemical moieties (e.g., alkyl, aryl, etc.).Nevertheless, such terms may also be used to convey correspondingmultivalent moieties under the appropriate structural circumstancesclear to those skilled in the art. For example, while an “alkyl” moietygenerally refers to a monovalent radical (e.g. CH₃—CH₂—), in certaincircumstances a bivalent linking moiety can be “alkyl,” in which casethose skilled in the art will understand the alkyl to be a divalentradical (e.g., —CH₂—CH₂—), which is equivalent to the term “alkylene.”(Similarly, in circumstances in which a divalent moiety is required andis stated as being “aryl,” those skilled in the art will understand thatthe term “aryl” refers to the corresponding divalent moiety, arylene.)All atoms are understood to have their normal number of valences forbond formation (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 forS, depending on the oxidation state of the S).

The term “amino” refers to —NH₂.

The term “acetyl” refers to “—C(O)CH₃.

As herein employed, the term “acyl” refers to an alkylcarbonyl orarylcarbonyl substituent wherein the alkyl and aryl portions are asdefined herein.

The term “alkyl” as employed herein refers to saturated straight andbranched chain aliphatic groups having from 1 to 12 carbon atoms. Assuch, “alkyl” encompasses C₁, C₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁and C₁₂ groups. Examples of alkyl groups include, without limitation,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,tert-butyl, pentyl, and hexyl.

The term “alkenyl” as used herein means an unsaturated straight orbranched chain aliphatic group with one or more carbon-carbon doublebonds, having from 2 to 12 carbon atoms. As such, “alkenyl” encompassesC₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ and C₁₂ groups. Examples ofalkenyl groups include, without limitation, ethenyl, propenyl, butenyl,pentenyl, and hexenyl.

The term “alkynyl” as used herein means an unsaturated straight orbranched chain aliphatic group with one or more carbon-carbon triplebonds, having from 2 to 12 carbon atoms. As such, “alkynyl” encompassesC₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ and C₁₂ groups. Examples ofalkynyl groups include, without limitation, ethynyl, propynyl, butynyl,pentynyl, and hexynyl.

An “alkylene,” “alkenylene,” or “alkynylene” group is an alkyl, alkenyl,or alkynyl group, as defined hereinabove, that is positioned between andserves to connect two other chemical groups. Examples of alkylene groupsinclude, without limitation, methylene, ethylene, propylene, andbutylene. Exemplary alkenylene groups include, without limitation,ethenylene, propenylene, and butenylene. Exemplary alkynylene groupsinclude, without limitation, ethynylene, propynylene, and butynylene.

The term “alkoxy” refers to -0C₁-C₆ alkyl.

The term “cycloalkyl” as employed herein is a saturated and partiallyunsaturated cyclic hydrocarbon group having 3 to 12 carbons. As such,“cycloalkyl” includes C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ and C₁₂cyclic hydrocarbon groups. Examples of cycloalkyl groups include,without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

The term “heteroalkyl” refers to an alkyl group, as defined hereinabove,wherein one or more carbon atoms in the chain are independently replacedO, S, or NRX, wherein Rx is hydrogen or C₁-C₃ alkyl. Examples ofheteroalkyl groups include methoxymethyl, methoxyethyl andmethoxypropyl.

An “aryl” group is a C₆-C₁₄ aromatic moiety comprising one to threearomatic rings. As such, “aryl” includes C₆, C₁₀, C₁₃, and C₁₄ cyclichydrocarbon groups. An exemplary aryl group is a C₆-C₁₀ aryl group.Particular aryl groups include, without limitation, phenyl, naphthyl,anthracenyl, and fluorenyl. An “aryl” group also includes fusedmulticyclic (e.g., bicyclic) ring systems in which one or more of thefused rings is non-aromatic, provided that at least one ring isaromatic, such as indenyl.

An “aralkyl” or “arylalkyl” group comprises an aryl group covalentlylinked to an alkyl group wherein the moiety is linked to another groupvia the alkyl moiety. An exemplary aralkyl group is—(C₁-C₆)alkyl(C₆-C₁₀)aryl, including, without limitation, benzyl,phenethyl, and naphthylmethyl. For example, an arC₁-C₃alkyl is an arylgroup covalently linked to a C₁-C₃ alkyl.

A “heterocyclyl” or “heterocyclic” group is a mono- or bicyclic (fusedor spiro) ring structure having from 3 to 12 atoms, (3, 4, 5, 6, 7, 8,9, 10, 11 or 12 atoms), for example 4 to 8 atoms, wherein one or morering atoms are independently —C(O)—, N, NR⁴, O, or S, and the remainderof the ring atoms are quaternary or carbonyl carbons. Examples ofheterocyclic groups include, without limitation, epoxy, oxiranyl,oxetanyl, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl,tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl,imidazolidinyl, thiazolidinyl, thiatanyl, dithianyl, trithianyl,azathianyl, oxathianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl,decahydroquinolinyl, piperidonyl, 4-piperidonyl, thiomorpholinyl,dimethyl-morpholinyl, and morpholinyl. Specifically excluded from thescope of this term are compounds having adjacent ring O and/or S atoms.

As used herein, “L-heterocyclyl” refers to a heterocyclyl groupcovalently linked to another group via an alkylene linker.

As used herein, the term “heteroaryl” refers to a group having 5 to 14ring atoms, preferably 5, 6, 10, 13 or 14 ring atoms; having 6, 10, or14 π electrons shared in a cyclic array; and having, in addition tocarbon atoms, from one to three heteroatoms that are each independentlyN, O, or S. Heteroaryl also includes fused multicyclic (e.g., bicyclic)ring systems in which one or more of the fused rings is non-aromatic,provided that at least one ring is aromatic and at least one ringcontains an N, O, or S ring atom. Examples of heteroaryl groups includeacridinyl, azocinyl, benzimidazolyl, benzofuranyl,benzo[d]oxazol-2(3H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one,benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl,benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl,benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl,chromenyl, cinnolinyl, furanyl, furazanyl, imidazolinyl, imidazolyl,1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl,isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl,isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl,octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl,oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole,pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl,2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl,quinoxalinyl, quinuclidinyl, tetrahydroisoquinolinyl,tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl,1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl,1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl,thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, andxanthenyl.

A “L-heteroaralkyl” or “L-heteroarylalkyl” group comprises a heteroarylgroup covalently linked to another group via an alkylene linker.Examples of heteroalkyl groups comprise a C₁-C₆ alkyl group and aheteroaryl group having 5, 6, 9, or 10 ring atoms. Examples ofheteroaralkyl groups include pyridylmethyl, pyridylethyl,pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl,thiazolylmethyl, thiazolylethyl, benzimidazolylmethyl,benzimidazolylethyl quinazolinylmethyl, quinolinylmethyl,quinolinylethyl, benzofuranylmethyl, indolinylethyl isoquinolinylmethyl,isoinodylmethyl, cinnolinylmethyl, and benzothiophenylethyl.Specifically excluded from the scope of this term are compounds havingadjacent ring O and/or S atoms.

An “arylene,” “heteroarylene,” or “heterocyclylene” group is a bivalentaryl, heteroaryl, or heterocyclyl group, respectively, as definedhereinabove, that is positioned between and serves to connect two otherchemical groups.

As employed herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl,heterocyclyl, urea, etc.) is described as “optionally substituted”without expressly stating the substituents it is meant that the groupoptionally has from one to four, preferably from one to three, morepreferably one or two, non-hydrogen substituents.

The term “halogen” or “halo” as employed herein refers to chlorine,bromine, fluorine, or iodine.

The term “haloalkyl” refers to an alkyl chain in which one or morehydrogens have been replaced by a halogen. Exemplary haloalkyls aretrifluoromethyl, difluoromethyl, flurochloromethyl, chloromethyl, andfluoromethyl.

The term “hydroxyalkyl” refers to -alkylene-OH.

EXAMPLE

The methods of the disclosure are illustrated further by the followingexamples, which is not to be construed as limiting the disclosure inscope or spirit to the specific procedures and compounds described inthem.

Study Design

The PRMT5 inhibitors of the disclosure demonstrate selective activity inMTAP-deleted cancers by binding to and further inhibiting PRMT5 whenbound to the intracellular metabolite MTA. As noted above, MTAP is anenzyme in the methionine salvage pathway and its deletion in cancercells leads to the accumulation of MTA in these cells. PRMT5 is anessential enzyme required for cell viability and, as such, the PRMT5inhibitors of the disclosure represent a novel approach to selectivelytreat MTAP-deleted cancers.

A single mutation will likely not cause cancer—most often, it ismultiple mutations that are responsible for developing cancer. Theinventors found the treatment of certain cancers with PRMT5 inhibitorsimproved with the use of combination therapies. Particularly, theinventors surprisingly found that a combination therapy of PRMT5inhibitor and KRAS^(G12C) inhibitor provides greater antitumor activitycompared to either inhibitor alone.

Study Procedure

Immunodeficient female nu/nu mice were implanted with 5×10⁶ LU99 lungcancer cells in 50% Matrigel. Tumors were measured using calipers untilthey reached approximately 150-200 mm³. Animals were randomized toreceive A) vehicle (0.5% methylcellulose (4000 cps)/0.2% Tween80 inwater), B) a PRMT5 inhibitor, C) KRAS^(G12C) inhibitor, or D) the PRMT5inhibitor and KRAS^(G12C) inhibitor, all administered orally (PO) for 21days. Tumor volume was measured twice a week (n=5/treatment group).Average tumor volume and standard error of the mean was calculated andplotted at each study day in GraphPad.

Example 1

This example was carried out according to the study procedure describedabove. The PRMT5 inhibitor was MRTX9768 administered at 100 mg/kg twicea day (BID). MRTX9768 is2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-3-fluoro-1-naphthonitrile,disclosed as Example 16-1 at p. 304 of the International patentpublication No. WO 2021/050915 A1, published 18 March 2021, incorporatedby reference in its entirety.

The KRAS^(G12C)inhibitor used in this example was MRTX849 administeredat 30 mg/kg once a day (QD). MRTX849 (adagrasib) is2-((S)-4-(7-(8-Chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-Amethoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile,disclosed in FIG. 1A of Hallin et al. 2020, Cancer Discov. 10(1): 54-71.

Results are provided in FIG. 1 and Table 1. The combination of MRTX9768and MRTX849 led to greater antitumor activity compared to eitherinhibitor alone in this KRAS^(G12C) and CDKN2A/MTAPDEL lung tumorxenograft LU99 model.

TABLE 1 Tumor Volume (mm³) Group Day 0 3 7 10 14 17 21 Vehicle Mean181.04 276.67 467.48 718.83 1040.93 1358.61 1951.06 (PO QD) SEM 18.8429.58 61.05 103.76 215.23 266.16 306.05 MRTX9768 Mean 183.59 265.26331.80 349.53 406.58 431.91 458.95 (100 mg/kg SEM 15.07 16.07 24.1425.13 43.52 51.50 60.11 PO BID) MRTX849 Mean 183.46 212.01 235.54 208.33238.62 253.40 302.46 (30 mg/kg SEM 15.02 24.43 31.54 30.73 50.71 64.2881.82 PO QD) MRTX9768 Mean 184.78 193.87 133.82 106.53 91.74 67.44 46.84(100 mg/kg SEM 15.41 23.01 12.32 9.73 9.18 9.54 10.26 PO BID) + MRTX849(30 mg/kg PO QD)

Example 2

This example was carried out substantially according to the studyprocedure described above. The PRMT5 inhibitor was MRTX7477,administered at 200 mg/kg BID. MRTX7477 is2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-1-naphthonitrile,disclosed as Example 4-147 at p. 226 of the International patentpublication No. WO 2021/050915 A1, published 18 March 2021, incorporatedby reference in its entirety. The KRAS^(G12C) inhibitor used in thisexample was the same as in Example 1, MRTX849, administered at 30 mg/kgQD.

Results are provided in FIG. 2 and Table 2. The combination of MRTX7477and MRTX849 led to greater antitumor activity compared to eitherinhibitor alone in this KRAS^(G12C) and CDKN2A/MTAPDEL lung tumorxenograft LU99 model.

TABLE 2 Tumor Volume (mm³) Group −2 1 5 8 12 15 19 21 Vehicle 126.3230.8 372.8 511.0 724.4 981.3 1357.6 1669.9 (PO QD) MRTX7477 129.4 214.5210.7 177.1 185.3 189.1 229.5 267.5 (200 mg/kg PO BID) MRTX849 130.1242.7 122.6 113.4 196.2 261.5 552.8 601.6 (30 mg · kg PO QD) MRTX7477131.0 224.8 99.8 60.8 60.7 45.6 46.7 64.4 (200 mg/kg PO BID) + MRTX849(30 mg/kg PO QD)

Example 3

The compound of the disclosure was evaluated in LU99 PRMT5-041 tumorxenograft model, and the results are provided in FIG. 3 and Table 3.This example was carried out substantially according to the studyprocedure described above, except with mice bearing LU99 PRMT5-041xenograft tumors. The PRMT5 inhibitor was MRTX1719, administered at 50mg/kg QD. MRTX1719 is(2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile,disclosed as Example 16-8 at p. 307 of the International patentpublication No. WO 2021/050915 Al, published 18 March 2021, incorporatedby reference in its entirety. The KRAS^(G12C) inhibitor used in thisexample was the same as in Example 1, MRTX849, administered at 30 mg/kgQD.

TABLE 3 Tumor Volume (mm³) Group Day 0 5 8 12 15 20 22 26 Vehicle Mean152 180 304 545 718 1140 1237 1548 (PO QD) SEM 12 19 52 100 133 193 162217 MRTX1719 Mean 153 135 145 164 163 167 176 203 (50 mg/kg SEM 13 18 2025 28 30 34 42 PO QD) MRTX849 Mean 154 142 117 87 81 84 103 171 (30mg/kg SEM 17 17 10 8 11 22 35 69 PO QD) MRTX1719 Mean 153 109 91 41 19 54 3 (50 mg/kg SEM 21 10 6 11 6 2 2 3 PO QD) + MRTX849 (30 mg/kg PO QD)

Example 4

The compound of the disclosure was evaluated in SW1573 PRMT5-044 tumorxenograft model, and the results are provided in FIG. 4 and Table 4.This example was carried out substantially according to the studyprocedure described above, except with mice bearing SW1573 PRMT5-044xenograft tumors. The PRMT5 inhibitor was MRTX1719, administered at 50mg/kg QD. The KRAS^(G12C)inhibitor used in this example was the same asin Example 1, MRTX849, administered at 100 mg/kg QD.

TABLE 4 Tumor Volume (mm³) Group Day 0 3 8 10 14 17 26 Vehicle Mean 145182 273 355 471 590 727 (PO QD) SEM 12 9 27 42 57 87 121 MRTX1719 Mean138 163 259 324 375 437 547 (50 mg/kg SEM 9 19 40 46 55 72 117 PO QD)MRTX849 Mean 139 150 188 236 285 320 361 (100 mg/kg SEM 9 14 19 28 30 5045 PO QD) MRTX1719 Mean 142 165 181 208 230 249 261 (50 mg/kg SEM 9 1721 34 42 52 61 PO QD) + MRTX849 (100 mg/kg PO QD)

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be incorporated within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated herein by referencefor all purposes.

What is claimed is:
 1. A method for treating cancer in a subject, themethod comprising: administering to the subject a therapeuticallyeffective amount of a Kirsten rat sarcoma viral oncogene homologglycine-to-cysteine (KRAS^(G12C)) inhibitor and a therapeuticallyeffective amount of a protein arginine N-methyl transferase 5 (PRMT5)inhibitor.
 2. The method of claim 1, wherein the cancer comprisesmethylthioadenosine phosphorylase (MTAP) gene homozygous deletion. 3.The method of claim, wherein the cancer comprises KRAS^(G12C) genemutation.
 4. The method of claim 2 , wherein the cancer further comprisea cyclin-dependent kinase inhibitor 2A (CDKN2A) gene homozygousdeletion.
 5. The method of claim 1, wherein the cancer is lung cancer,pancreatic cancer, colon cancer, head and neck cancer, esophagealcancer, or melanoma.
 6. The method of claim 1, wherein the cancer islung cancer, such as non-small cell lung cancer (NSCLC).
 7. The methodof claim 1, wherein the cancer is pancreatic or colon cancer.
 8. Themethod of claim 1, wherein the KRAS^(G12C) inhibitor is selected fromadagrasib, sotorasib, JNJ-74699157, GDC-6036, LY3499446, JDQ443, D-1553,and combinations thereof.
 9. The method of claim 1, wherein theKRAS^(G12C) inhibitor is adagrasib.
 10. The method of claim 1, whereinthe KRAS^(G12C) inhibitor is sotorasib.
 11. The method of claim 1,wherein the PRMT5 inhibitor is a methylthioadenosine (MTA)-cooperativePRMT5 inhibitor.
 12. The method of claim 1, wherein the PRMT5 inhibitoris a compound of Formula IIA, IIB or IIC:

or a pharmaceutically acceptable salt thereof, wherein: A is CR⁹ or N; Dis (C(R⁹)₂)₁₋₂—NH₂,

or D is

where the methylene is bonded to E where E is C; E is C, CR⁹ or N; eachL is independently a bond or C₁-C₃ alkylene; W is CR⁹ or N; each X isindependently a bond, O, S, —NR⁴— or —NR⁴C(O)—; each Z is independentlya bond, —SO—, —SO₂—, —CH(OH)— or —C(O)—; each R² is independentlyhydroxy, halogen, cyano, cyanomethyl, —(NR⁴)₂, hydroxyalkyl, alkoxy,—SO₂C₁-C₃alkyl, —X-arC₁-C₃alkyl, heteroalkyl, C₂-C₄ alkynyl,—X-haloalkyl, —X-C₁-C₅ alkyl, —Z-C₁-C₅ alkyl, heterocyclyl,—X—L-cycloalkyl, —Z-cycloalkyl, —X-aryl, —Z-aryl, or -X-heteroaryl,wherein the heterocyclyl, the cycloalkyl, the aryl and the heteroarylare optionally substituted with one or more R⁵; each R⁴ is independentlyhydrogen or C₁-C₃ alkyl; each R⁵ is independently cyano, oxo, halogen,C₁-C₃ alkyl, hydroxyalkyl, hydroxy, alkoxy, alkoxy-C₁-C₃ alkyl,—X-haloalkyl, —Z-cycloalkyl, —X-arC₁-C₃alkyl, —X-arC₁-C₃alkylsubstituted with cyano, —X—L-cycloalkyl optionally substituted withC₁-C₃ alkyl or oxo, —X—L-heteroaryl optionally substituted with one ormore C₁-C₃ alkyl or oxo, —X—L-heterocyclyl optionally substituted withone or more C₁-C₃ alkyl or oxo, or —X-aryl; R⁶ is hydrogen, halogen,C₁-C₃ alkyl, haloalkyl, hydroxy, alkoxy, C₁-C₃ alkyl-alkoxy, N(R⁹)₂,NR⁹C(O)R⁹, C(O)R⁹, oxetane and THF; R⁷ is H or C₁-C₃ alkyl optionallysubstituted with one or more halogen; R⁸ is H or C₁-C₃ alkyl; and eachR⁹ is independently H or C₁-C₃ alkyl, halogen or haloalkyl.
 13. Themethod of claim 1, wherein the PRMT5 inhibitor is a compound of FormulaIIIA:

or a pharmaceutically acceptable salt thereof, wherein A is CR⁹ or N; Dis —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl; R² is

where R⁵⁶ is hydrogen, fluoro, chloro, or methyl, G, Q, J and U areindependently selected from C(H), C(R⁵), and N, provided only one or twoof G, Q, J, and U can be N; each R⁵ is independently hydroxy, halogen,C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, C₃-C₆cycloalkoxy, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃alkoxyC₁-C₃ alkyl; R⁶ is hydrogen, halogen, C₁-C₆ alkyl, hydroxy, C₁-C₆alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)-C₁-C₃haloalkyl, or —NR¹⁵(CO)R¹⁶, where R¹⁵ is hydrogen or methyl, and R¹⁶ isC₁-C₃ alkyl; and R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.
 14. The method ofclaim 13, wherein the PRMT5 inhibitor is:

or a pharmaceutically acceptable salt thereof.
 15. The method of claim1, wherein the PRMT5 inhibitor is a compound of Formula IIIB:

or a pharmaceutically acceptable salt thereof, wherein A is CR⁹ or N; Dis —CH₂—NH₂,

W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl; R⁵¹ is hydrogen, fluoro,chloro, or methyl, or R⁵¹ and R⁵² together with atoms to which they areattached form a C₄-C₆ heterocycloalkyl (e.g, hydrofuranyl); R⁵² isfluoro, chloro, or methyl, or R⁵² and R⁵³ together with atoms to whichthey are attached form a phenyl; R⁵³ is hydrogen, fluoro, chloro, ormethyl; R⁵⁴ is hydrogen, halogen, C₁-C₃ alkyl, or C₁-C₃ alkoxy; L⁵ is—O— or —CH₂—; R⁶ is hydrogen, halogen, C₁-C₆ alkyl, hydroxy, C₁-C₆alkoxy, C₁-C₀₃ alkoxyC₁-C₃ alkyl, C₃-C₆ heterocycloalkyl, —C(O)—C₁-C₃haloalkyl, or —NR¹⁵(CO)R¹⁶, where R¹⁵ is hydrogen or methyl, and R¹⁶ isC₁-C₃ alkyl; R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl.
 16. The method ofclaim 15, wherein: A is —CH or —CCH₃; D is —CH₂—NH₂; W is —CH, —CCH₃, orN; R⁵¹, R⁵², R⁵³, and R⁵⁴ are each independently selected from hydrogen,fluoro, chloro, or methyl; L⁵ is —O—; R⁶ is hydrogen, fluoro, chloro, ormethyl; and R⁷ is C₁-C₂ alkyl or C₁-C₂ haloalkyl.
 17. The method ofclaim 15, wherein: A and W are —CH; D is —CH₂—NH₂; R⁵¹, R⁵², and R⁵³ areeach independently selected from hydrogen, fluoro, chloro, and methyl;R⁵⁴ is hydrogen; L⁵ is —O—; R⁶ is hydrogen; and R⁷ is methyl.
 18. Themethod of claim 15, wherein: A and W are —CH; D is —CH₂—NH₂; R⁵¹ and R⁵²are each independently selected from fluoro, chloro, and methyl; R⁵³ andR⁵⁴ are hydrogen; L⁵ is —O—; R⁶ is hydrogen; and R⁷ is methyl.
 19. Themethod of claim 15, wherein the PRMT5 inhibitor is:

or a pharmaceutically acceptable salt thereof.
 20. The method of claim15, wherein the PRMT5 inhibitor is:

or a pharmaceutically acceptable salt thereof.
 21. The method of claim1, wherein the PRMT5 inhibitor is MRTX1719 or a pharmaceuticallyacceptable salt thereof, and the KRAS^(G12C)inhibitor is adagrasib. 22.The method of claim 1, wherein the PRMT5 inhibitor is MRTX1719 or apharmaceutically acceptable salt thereof, and the KRAS^(G12C) inhibitoris sotorasib.
 23. The method of claim 1, wherein the PRMT5 inhibitor isa compound of Formula IIIC:

or a pharmaceutically acceptable salt thereof, wherein A is CR⁹ or N;

D is —CH₂—NH₂, W is CR⁹ or N, where R⁹ is H or C₁-C₃ alkyl; G, Q, J andU are independently selected from C(H), C(R⁵), and N, provided only oneor two of G, Q, J, and U can be N; each R⁵ is independently hydroxy,halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy,C₃-C₆ cycloalkoxy, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, or C₁-C₃alkoxyC₁-C₃ alkyl; R⁶ is hydrogen, halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, hydroxy, C₁-C₆ alkoxy, C₁-C₃ alkoxyC₁-C₃ alkyl, C₃-C₆heterocycloalkyl, —C(O)—C₁-C₃ haloalkyl, —N(R⁹)₂, or —NR¹⁵(CO)R¹⁶, whereeach R⁹ is independently H or C₁-C₃ alkyl, R¹⁵ is hydrogen or methyl,and R¹⁶ is C₁-C₃ alkyl; and R⁷ is C₁-C₃ alkyl or C₁-C₃ haloalkyl. 24.The method of claim 23, wherein the PRMT5 inhibitor is:

or a pharmaceutically acceptable salt thereof.
 25. The method of claim1, wherein the therapeutically effective amount of the PRMT5 inhibitoris in the range of about 0.01 to 300 mg/kg per day.
 26. The method ofclaim 1, wherein the therapeutically effective amount of the PRMT5inhibitor is in the range of about 0.1 to 100 mg/kg per day.
 27. Themethod of claim 1, wherein the therapeutically effective amount of thePRMT5 inhibitor is less than 1% of, e.g., less than 10%, or less than25%, or less than 50% of the clinically-established therapeutic amount.28. The method of claim 1, wherein the therapeutically effective amountof the KRAS^(G12C) inhibitor is in the range of about 0.01 to 300 mg/kgper day.
 29. The method of claim 1, wherein the therapeuticallyeffective amount of the KRAS^(G12C) inhibitor is in the range of about0.1 to 100 mg/kg per day.
 30. The method of claim 1, wherein thetherapeutically effective amount of the KRAS^(G12C) inhibitor is lessthan 1% of, e.g., less than 10%, or less than 25%, or less than 50% ofthe clinically-established therapeutic amount.
 31. The method of claim1, wherein the KRAS^(G12C) inhibitor and the PRMT5 inhibitor areadministered sequentially.
 32. The method of claim 1, wherein theKRAS^(G12C) inhibitor and the PRMT5 inhibitor are administeredsimultaneously.
 33. The method of claim 1, wherein the subjectpreviously received or completed a first-line chemotherapy.
 34. Themethod of claim 33, wherein the first-line chemotherapy is platinum-and/or taxane-based chemotherapy.